Single Molecule Analysis of MAGUK Structure and Ligand Binding

MAGUK 结构和配体结合的单分子分析

基本信息

批准号：
10360516
负责人：
Mark E Bowen
金额：
$ 44.49万
依托单位：
STATE UNIVERSITY NEW YORK STONY BROOK
依托单位国家：
美国
项目类别：
财政年份：
2008
资助国家：
美国
起止时间：
2008-01-10 至 2025-03-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10360516
关键词：
Active Sites Address Affect Affinity Allosteric Regulation Award Binding Brain Clinical Cytoplasmic Tail Data Deposition Development Disease Epilepsy Event Family Filament Future Genetic Glutamate Receptor Glutamates In Vitro Individual Kinetics Knowledge Learning Length Ligand Binding Ligands Link Lipids Liquid substance Location Mediating Membrane Memory Mental disorders Methods Modification Molecular Molecular Conformation Mutation Nerve Degeneration Neurosciences Neurotransmitter Receptor Oils Organelles Outcome Peptides Phase Phase Transition Phosphorylation Phosphotransferases Physiological Play Post-Translational Protein Processing Post-Translational Regulation Protein Isoforms Protein Kinase Proteins Proteomics Publishing Reaction Receptor Signaling Role Scaffolding Protein Schizophrenia Sequence Homology Signal Transduction Signaling Protein Site Specificity Stroke Structural Models Structure Synapses Synaptic plasticity System Testing Translations Water Work autism spectrum disorder druggable target glutamatergic signaling in vivo interest membrane activity membrane-associated guanylate kinase nervous system disorder neuropsychiatric disorder neurotransmission novel palmitoylation particle post stroke receptor receptor binding reconstitution recruit scaffold single molecule

项目摘要

This proposal focuses on post-synaptic glutamate signaling, which mediates excitatory neurotransmission. Glutamate receptor signaling is organized by the membrane-associated guanylate kinase (MAGuK) family of scaffold proteins. Scaffolds determine the outcome of signal transduction by controlling the location of neurotransmitter receptors and connecting them to downstream effectors. In particular, two homologous MAGuKs (PSD-95 and PSD-93) play opposing roles in synaptic plasticity yet the basis for their differential activity remains a mystery. A key factor in synaptic plasticity is post-translation modification (PTM) of the MAGuK proteins, which have been missing in most studies that have been published to date. This proposal investigates the posttranslational regulation of PSD-95 and PSD-93. Aim 1 will test the hypothesis that phosphorylation alters the structure and activity of MAGuKs. We propose that a difference in the number and location of PTM sites elicits activity differences. Aim 2 will test the hypothesis that palmitoylation, a lipid modification essential for synaptic targeting, alters MAGuK activity and coverts them into the filament observed at synapses. Studies to date focused on soluble proteins but palmitoylation is indispensable for activity. Aim 3 will test the hypothesis that phase transitions of MAGuKs regulate availability for receptor binding. SynGAP, an essential synaptic protein, induces to liquid phase separation of PSD-95. This can generate a dynamic “membrane-less organelle”. We will investigate the functional effects of phase separation on receptor binding and structure. We possess the only working reconstitution of scaffold interactions in the post-synapse and provided the only kinetic description of MAGuK scaffolding activity. We will use our unique expertise to address how PTM changes the activity of MAGuKs. We have pioneered novel methods for structural refinement of proteins containing intrinsic disorder and generated the only structural model of a full-length scaffold protein to date. We will use these approaches to provide a mechanistic understanding of how MAGuK structure affects scaffolding. Our published work suggests that proteins residing together on PSD-95 have “higher-order” interactions driven by the enforced proximity. Describing the molecular events in excitatory signaling is a fundamental challenge in neuroscience with direct relevance to brain development, memory and learning, and many neurological and neuropsychiatric disorders.

该提案重点关注突触后谷氨酸信号传导，它介导兴奋性谷氨酸受体信号传导是由膜相关组织的。鸟苷酸激酶 (MAGuK) 支架蛋白家族决定了结果。通过控制神经递质受体的位置和连接来进行信号转导特别是，两个同源的 MAGuK（PSD-95 和 PSD-93）。在突触可塑性中发挥相反的作用，但它们差异活动的基础仍然是突触可塑性的一个关键因素是 MAGuK 的翻译后修饰 (PTM)。迄今为止已发表的大多数研究中都缺少蛋白质。该提案研究了 PSD-95 和 PSD-93 的翻译后调控。图 1 将检验磷酸化改变 MAGuK 的结构和活性的假设。认为 PTM 位点数量和位置的差异会引起活性差异。目标 2 将检验以下假设：棕榈酰化（突触必需的脂质修饰）靶向、改变 MAGuK 活性并将其转化为在突触处观察到的细丝。迄今为止的研究主要集中在可溶性蛋白质上，但棕榈酰化对于 Aim 的活性是必不可少的。 3 将检验 MAGuK 的相变调节受体可用性的假设 SynGAP 是一种重要的突触蛋白，可诱导 PSD-95 的液相分离。这可以产生动态的“无膜细胞器”，我们将研究其功能。相分离对受体结合和结构的影响。我们拥有突触后支架相互作用的唯一有效重建并提供了 MAGuK 脚手架活动的唯一动力学描述，我们将使用我们独特的。我们开创了新颖的解决方案，以解决 PTM 如何改变 MAGuK 的活动。对含有内在无序的蛋白质进行结构精炼的方法，并产生了唯一的迄今为止，我们将使用这些方法来构建全长支架蛋白的结构模型。提供对 MAGuK 结构如何影响脚手架的机械理解。研究表明，PSD-95 上共同存在的蛋白质具有“高阶”相互作用驱动通过强制接近来描述兴奋信号传导中的分子事件是一个神经科学中与大脑发育、记忆和记忆直接相关的基本挑战学习以及许多神经系统和神经精神疾病。

项目成果

期刊论文数量（25）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Domain orientation in the N-Terminal PDZ tandem from PSD-95 is maintained in the full-length protein.

DOI：
10.1016/j.str.2011.02.017
发表时间：
2011-06-08
期刊：
STRUCTURE
影响因子：
5.7
作者：
McCann, James J.;Zheng, Liqiang;Chiantia, Salvatore;Bowen, Mark E.
通讯作者：
Bowen, Mark E.

Integrative structural dynamics probing of the conformational heterogeneity in synaptosomal-associated protein 25

突触体相关蛋白 25 构象异质性的综合结构动力学探测

DOI：
10.1016/j.xcrp.2021.100616
发表时间：
2021
期刊：
Cell Reports Physical Science
影响因子：
8.9
作者：
Saikia, Nabanita;Yanez-Orozco, Inna S.;Qiu, Ruoyi;Hao, Pengyu;Milikisiyants, Sergey;Ou, Erkang;Hamilton, George L.;Weninger, Keith R.;Smirnova, Tatyana I.;Sanabria, Hugo
通讯作者：
Sanabria, Hugo

Structure and conformational dynamics of Clostridioides difficile toxin A.

DOI：
10.26508/lsa.202201383
发表时间：
2022-06
期刊：
Life science alliance
影响因子：
4.4
作者：
Chen B;Basak S;Chen P;Zhang C;Perry K;Tian S;Yu C;Dong M;Huang L;Bowen ME;Jin R
通讯作者：
Jin R

Beyond the random coil: stochastic conformational switching in intrinsically disordered proteins.

DOI：
10.1016/j.str.2011.01.011
发表时间：
2011-04-13
期刊：
STRUCTURE
影响因子：
5.7
作者：
Choi, Ucheor B.;McCann, James J.;Weninger, Keith R.;Bowen, Mark E.
通讯作者：
Bowen, Mark E.

Divergent roles of a peripheral transmembrane segment in AMPA and NMDA receptors.

DOI：
10.1085/jgp.201711762
发表时间：
2017-06-05
期刊：
The Journal of general physiology
影响因子：
0
作者：
Amin JB;Salussolia CL;Chan K;Regan MC;Dai J;Zhou HX;Furukawa H;Bowen ME;Wollmuth LP
通讯作者：
Wollmuth LP